1.Identification and functional characterization of a new flavonoid glycosyltransferase from Rheum palmatum.
Shiwen ZHANG ; Jianzhen ZOU ; Zitong HAO ; Mengqi GAO ; Gang ZHANG ; Mengmeng LIU
Chinese Herbal Medicines 2025;17(2):307-314
OBJECTIVE:
To characterize a glycosyltransferase (RpUGT1) from Rheum palmatum and investigate its specificity toward flavonoid compounds.
METHODS:
The RpUGT1 was expressed in Escherichia coli and screened for catalytic activity against a range of flavonoid substrates using a high-throughput HPLC assay method. Mass spectrometry (MS) and nuclear magnetic resonance (NMR) were used to determine the structure of the product. Homology modeling, molecular docking analyses and site-directed mutagenesis studies were conducted to identify key residues responsible for its function.
RESULTS:
The recombinant RpUGT1 protein exhibited catalytic activity towards various flavonoids. Notably, RpUGT1 catalyzed the glycosylation of isorhamnetin to form 3-O-glucoside and kaempferol to form 7-O-glucoside, utilizing uridine diphosphate (UDP) glucose as the sugar donor. The homology modeling and molecular docking analyses identified key residues responsible for its activity. Subsequent site-directed mutagenesis studies highlighted the crucial role of K307 in catalysis.
CONCLUSION
These discoveries offer valuable perspectives on the role of the UGT family and establish a groundwork for forthcoming research on the synthesis of flavonoids in plants.
2.Mechanism of protection of motor neurons in spinal cord anterior horn of SNI rats by acellular nerve allografts via the Bcl-2/Cyt-C/Apaf-1 signalling pathway
Mengyuan Zheng ; Zitong Hao ; Qinghua Zhu ; Zhuangzhuang Tian ; Xingda Guo ; Yuhe Zheng ; Cheng Li ; Xiumei Fu
Acta Universitatis Medicinalis Anhui 2025;60(11):2035-2042
Objective:
To investigate the protective effects and mechanisms of acellular nerve allografts (ANA) on motor neurons in the spinal cord anterior horn of sciatic nerve injury ( SNI) rats .
Methods:
SPF grade male SD rats were randomly divided into normal , model , ANA-bridged (bridge group) , and autologous nerve transplantation groups (autograft group) , with 6 rats in each group . The SNI rat model was established using the right sciatic nerve clamp method for 10 mm . In the bridge group , the ANA was bridged to the two severed ends of the injured sciatic nerve , and in the autograft group , the autologous nerves were flipped head to tail and then bridged to the two se- vered ends . A spectrophotometer was applied to determine the DNA content in normal nerves and ANA . The foot- print test was used to determine the sciatic nerve function index (SFI) of the rats in each group , the wet weight ra- tio of the anterior tibialis muscle was calculated . The morphology and structure of the anterior horn motor neurons of the spinal cord of each group were observed by HE staining. The immunofluorescence and Western blot were used to detect Apaf-1 , Caspase-3 , Bcl-2 , Bax , and Cyt-C proteins expression in the L4-6 segment of the spinal cord .
Results:
The DNA content in the ANA prepared in this study was significantly lower than that in normal nerves (P < 0. 05) . Compared with the normal group , the SFI and wet weight ratio of the anterior tibialis muscle were re- duced in the model group (P < 0. 001) ; compared with the model group , both SFI and wet weight ratio of the ante- rior tibialis muscle significantly increased in the bridge group and the autografts group ( P < 0. 05 , P < 0. 001) , and the SFI and wet weight ratio of the anterior tibialis muscle in the autograft group were higher than those in the bridge group (P < 0. 001 , P < 0. 01) . The results of HE staining showed that the motor neurons in the anterior horn of the spinal cord of the normal group were structurally intact and had clear cytosolic boundaries; the neurons in the model group were lysed and necrotic , with blurred cytosolic boundaries; the neurons in the bridge group were less lysed and necrotic , but the nuclear translocation phenomenon could still be seen; the neurons in the autograft group were morphologically and structurally intact with clear cytosolic boundaries . Compared with the normal group , the expression of Apaf-1 , Caspase-3 , Bax and Cyt-C proteins significantly increased in the model group (P < 0. 001 , P < 0. 01 , P < 0. 01 , P < 0. 05) . Compared with the model group , the expression of Apaf-1 , Caspase- 3 , Bax , and Cyt-C proteins significantly decreased (P < 0. 001 , P < 0. 05 , P < 0. 05 , P < 0. 05) ; but the expres- sion of Bcl-2 protein significantly increased in the bridge group and the autograft group (P < 0. 05) . The expression of Apaf-1 , Caspase-3 , Bax and Cyt-C proteins in the autografts group was lower than that in the bridge group (P < 0. 001 , P < 0. 05 , P < 0. 05 , P < 0. 05) .
Conclusion
ANA can exert a protective effect on motor neurons in the anterior horn of the spinal cord of SNI rats by improving the morphology and structure of neurons , increasing the ex- pression of Bcl-2 protein , but decreasing the expression of Cyt-C , Bax , Caspase-3 , and Apaf-1 proteins in the spi- nal cord . The mechanism of ANA may be related to the Bcl-2/Cyt-C/Apaf-1-mediated mitochondrial apoptosis sig- naling pathway .
3.Alcohol Abstinence and Accelerated Biological Aging Among Middle-Aged and Older Adults:Evidence From the UK Biobank
Hongxiang CHEN ; Jiajie CAI ; Jun WEI ; Hongmei ZHANG ; Yi XIANG ; Zitong HUANG ; Hao XU ; Xiong XIAO ; Xing ZHAO
Journal of Sichuan University (Medical Sciences) 2024;55(2):353-359
Objective To investigate the longitudinal association between alcohol abstinence and accelerated biological aging among middle-aged and older adults and to explore the potential effect modifiers influencing the association.Methods Utilizing the clinico-biochemical and anthropometric data from the baseline and first repeat survey of the UK Biobank(UKB),we employed the Klemera and Doubal method(KDM)to construct the biological age(BA)and calculate BA acceleration.Change analysis based on multivariate linear regression models was employed to explore the association between changes in alcohol abstinence and changes in BA acceleration.Age,sex,smoking status,tea and coffee consumption,and body mass index were considered as the stratification factors for conducting stratified analysis.Results A total of 5 412 participants were included.Short-term alcohol abstinence(β=1.00,95%confidence interval[CI]:0.15-1.86)was found to accelerate biological aging when compared to consistent never drinking,while long-term abstinence(β=-0.20,95%CI:-1.12-0.71)did not result in a significant acceleration of biological aging.Body mass index may be a potential effect modifier.Conclusion Short-term alcohol abstinence was associated with accelerated biological aging,but the effect gradually diminishes over extended periods of abstinence.
4.开放科学(资源服务)标识码(OSID) Effect of Tanshinone ⅡA on abnormal integration of granule cells in the dentate gyrus of mice with chronic epilepsy
Xinli XIAO ; Zitong QIN ; Kaixiang REN ; Yiqiu ZHENG ; Hao PENG ; Qiqi QIANG ; Chutong ZHANG ; Zhuoyuan YANG ; Qi LEI ; Jianxin LIU
Journal of Xi'an Jiaotong University(Medical Sciences) 2023;44(4):634-643
【Objective】 To clarify the role and molecular mechanism of Tanshinone ⅡA (TanⅡA) in the pathological integration of granule cells in the dentate gyrus (DG) by using the mouse model of temporal lobe epilepsy (TLE). 【Methods】 Status epilepticus (SE) was induced in the mice with pilocarpine and treated with TanⅡA 5 mg/kg. After two months, Morris water maze was used to examine the spatial learning and memory ability and video surveillance was used to monitor spontaneous seizures. The DG was removed for staining of Timm, Prox-1, DCX and SynⅠ. PTEN, p-AKT, and p-S6 expressions were observed by Western blotting. 【Results】 TanⅡA decreased Timm score, SynⅠ, PSD-95 and pS6 levels, and increased the level of PTEN in the DG, and attenuated the formation of mossy fiber sproutings and basal dendrites of the granule cells. Video surveillance showed that TanⅡA reduced the frequency of Racine’ grade 5 seizures. 【Conclusion】 TanⅡA can effectively attenuate the abnormal integration of the granule cells in the DG by regulating PTEN/AKT/mTOR pathway and thus plays an anti-epileptic role.
5.Evaluation of temporomandibular joint space and condylar morphology in patients with anterior open-bite based on cone-beam CT
LAI Zhanwen ; HU Ziyang ; PAN Xiao ; HAO Yanqing ; LIN Zitong
Journal of Prevention and Treatment for Stomatological Diseases 2021;29(7):468-473
Objective:
To investigate the difference of the temporomandibular joint between patients with anterior open-bite and normal overbite with cone beam CT (CBCT).
Methods :
Fifty-four patients with anterior open bites and 54 patients with normal overbites were selected from the Department of Orthodontics, Nanjing Stomatological Hospital, Medical School of Nanjing University from June 2014 to August 2020. Sagittal and coronal images of the temporomandibular joint were reconstructed with multiplanar reconstruction techique. The Kamelchuk method was used to measure the superior, posterior and anterior space of the temporomandibular joint, and the condylar morphology was divided into two types: normal and abnormal. The joint space and condylar morphology of the anterior open-bite group and the normal overbite group were statistically analyzed. The anterior open-bite group was divided into 3 subgroups: ① Ⅰ° open-bite (open bite distance < 3 mm), ② Ⅱ° open-bite (open bite distance ≥ 3 mm and ≤ 5 mm) and ③ Ⅲ° open-bite (open bite distance > 5 mm). The difference of overbite spaces of the temporomandibular joint was compared among these three subgroups.
Results:
Compared to the normal group, no significant differences were found for the anterior and superior space of the temporomandibular joint in the anterior open-bite group (P > 0.05), but the posterior space increased significantly (P < 0.01). A total of 52.8% of patients in the anterior open-bite group had abnormal condyles, whereas 21.3% of patients in the normal group, overbite significant differences was found between the two groups (P < 0.01). Compared with patients with Ⅰ° and Ⅱ° openbite, the condyle of patients with III° open bites was more forward in the fossa (P < 0.05).
Conclusion
The position of the condyle in the fossa of patients with anterior open bites is more forward, and abnormal condylar bone is more common found.
6.Preliminary observation of the expressions of SOCS-1 and SOCS-3 in myocardium of patients with sudden cardiac death
Liang CHEN ; Zhongfu MA ; Hao TANG ; Yanbing LIANG ; Zhibin CHEN ; Zhenyu LI ; Zitong HUANG ; Longyuan JIANG
Chinese Journal of Emergency Medicine 2010;19(3):277-280
Objective To investigate the expressions and clinical significanees of suppressors of cytokine signaling-1 (SOCS-1) and SOCS-3 in myocardium of patients with sudden cardiac death (SCD). Method This study included myocardial autopsy specimens of 24 patients admitted between 2005 and 2006. Of them, 9 cases had the findings of autopsy examination consistent with coronary atberosclerosis (non-myocardial infarction) leading to SCD (non-MI group), 7 patients died of acute myocardial infarction (MI group) and 8 patients died of traffic accidents and trauma The expressions of SOCS-1 mRNA and SOCS-3 mRNA in the myocardium of non-MI and con-trol group were detected by using RT-PCR. The levels of SOCS-1 protein and SOCS-3 protein were detected by us-ing immunohistochemistry. Statistical analysis were performed by using SPSS version 13.0 software and the data were processed with ANOVA test. Results The expressions of SOCS-1 mRNA and SOCS-3 mRNA in non-MI and MI groups were were significantly higher than those in control group (0. 788±0. 101) and (0. 741±0.111) vs.(0.436±0.044) (P <0.01); (0.841±0.092) and (0.776±0.070) vs.(0.454±0.076), P <0.01, re-spectively). The antibody-positive cells of SOCS-1 protein in myocardium of non-MI group and MI group were significantly higher than those in myoeardium of control group (320.00±48.48) and (347.14±70.88) vs.(42.50±10.35) (P < 0.01), respectively. The antibody-positive cells of SOCS-3 protein in myoeardium of non-MI group and MI group were significantly higher than those in myocardium of control group (381.11±59.25) vs.(40.00±10.69), (P < 0.01)and (332.86±111.91) vs. (40.00±10.69), (P =0.001). Conclusions The expressions of SOCS rnRNA and SOCS-3 mRNA in myoeardium of patients with SCD from coronary diseases are significantly increased contributing to the pathogenesis of SCD.


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